CBMN, UMR 5248 CNRS, Université de Bordeaux, Bat B14, allée Geoffroy St. Hilaire, Pessac, France.
Institute of Electronics, Microelectronics and Nanotechnology (IEMN),UMR-CNRS 8520, Université Lille 1, Cité Scientifique, 59655 Villeneuve d'Ascq, France; Taras Shevchenko University, 60 Vladimirskaya Str., Kiev, Ukraine.
Anal Chim Acta. 2015 May 11;873:71-9. doi: 10.1016/j.aca.2015.02.060. Epub 2015 Feb 24.
Carbohydrate-modified interfaces have been shown to be valuable tools for the study of protein-glycan recognition events. Label-free approache such as plasmonic based techniques are particularly attractive. This paper describes a new analytical platform for the sensitive and selective screening of carbohydrate-lectin interactions using plasmon waveguide resonance. Planar optical waveguides (POW), consisting of glass prisms coated with silver (50 nm) and silica (460 nm) layers were derivatized with mannose or lactose moieties. The specific association of the resulting interface with selected lectins was assessed by following the changes in its plasmonic response. The immobilization strategy investigated in this work is based on the formation of a covalent bond between propargyl-functionalized glycans and surface-linked azide groups via a Cu(I) "click" chemistry. Optimization of the surface architecture through the introduction of an oligo(ethylene glycol) spacer between the plasmonic surface and the glycan ligands provided an interface which allowed screening of glycan-lectin interactions in a highly selective manner. The limit of detection (LOD) of this method for this particular application was found to be in the subnanomolar range (0.5 nM), showing it to constitute a promising analytical platform for future development and use in a pharmaceutical or biomedical setting.
糖基化修饰界面已被证明是研究蛋白-聚糖识别事件的有用工具。基于等离子体的无标记方法特别有吸引力。本文描述了一种使用等离子体波导共振进行敏感和选择性筛选糖-凝集素相互作用的新分析平台。平面光波导 (POW) 由涂有银 (50nm) 和二氧化硅 (460nm) 层的玻璃棱镜组成,用甘露糖或乳糖部分进行衍生化。通过监测等离子体响应的变化,评估了所得界面与选定凝集素的特异性结合。本工作中研究的固定化策略基于炔丙基功能化聚糖与通过 Cu(I) “点击”化学连接的表面连接叠氮基团之间形成共价键。通过在等离子体表面和糖配体之间引入聚乙二醇 (PEG) 间隔物来优化表面结构,提供了一种允许以高度选择性方式筛选糖-凝集素相互作用的界面。该方法对于这种特定应用的检测限 (LOD) 为亚纳摩尔范围 (0.5 nM),表明它构成了未来在制药或生物医学环境中开发和使用的有前途的分析平台。